Neural induction of human ESCs to NESCs.

<p>A) Schematic representation of the protocol used to differentiate the ESC line H9 to NESCs, entailing the formation of embryoid bodies, followed by neural rosette formation and mechanical isolation. Bar scale 100 μm. B) NESCs cultured in EGF and FGF2 containing medium staining positive for SOX2 in the nucleus (red) and NESTIN in the cytoplasm (green), bar scale 100 μm. NESCs showed a tendency to distribute in rosettes-like structures (white arrow), with the nuclei located at the center and the cytoplasms at the periphery. C) Expression level of pluripotency and neural factors in the three independent cultures of ESCs and NESCs as obtained from the microarray analysis. Genes marked by asterisks were significantly differentially expressed (p ≤0.001***, p ≤0.0001 ****, unpaired t test with Bonferroni correction).